乙酰化和非乙酰化檢測試劑盒的差異:

目前市場上的 cAMP 和 cGMP 檢測試劑盒的一個關鍵步驟是將乙酸酐含與cAMP或 cGMP的樣品一起孵育，以形成乙酰化的 cAMP 或 cGMP 衍生物。然后抗體與乙酰化 cAMP 或 cGMP 中間體結合,以提高靈敏度。眾所周知，醋酸酐是一種毒性很強的強乙酰化劑, 長期接觸會損害科研人員的健康。乙酸酐還可以乙酰化ATP、GTP或其它的芳環。乙酰化產物也可以被抗體識別，從而降低了靈敏性。
Product Highlight

• Based on the unique mouse monoclonal anti-cAMP antibody
• The only assay kit on the market with monoclonal anti-cAMP antibody (all other kits are based on polyclonal anti-cAMP antibodies)
• The monoclonal anti-cAMP antibody displays much higher selectivity over cGMP and ATP than polyclonal anti-cAMP antibody
• Avoids the batch-to-batch variations associated with polyclonal antibody productions from animals, thus providing the reproducibility

 
Principle Outline
NewEast Biosciences cAMP ELISA Kit is a competitive immunoassay to measure the cAMP levels, either from cell extracts or from in vitro adenylyl cyclase assays. Briefly, multi-well plates are coated with goat-anti-mouse serum. cAMP in cell extracts or in in vitro adenylyl cyclase assays will competitively bind to the monoclonal anti-cAMP antibody in the presence of fixed amounts of cAMP-conjugated horse-radish peroxidase or alkaline phosphatase. Known amounts of cAMP are used as standards to generate the calculation curve. After a short incubation, the excess reagents are washed away and substrate is added. The multiwell plates are then read on a microplate reader at 450 nm or 405 nm. The intensity of the yellow color is inversely proportional to the concentration of cAMP in samples. The measured optical density is used to calculate the concentration of cAMP in samples based on the curve from the cAMP standards.

NewEast Biosciences 花了兩年時間篩選大量的單克隆抗體，從中篩出識別裸 cAMP 或 cGMP抗體，而不識別 ATP 和 GTP。因此，本公司試劑盒省去了使用有毒乙酸酐的乙酰化步驟, 提高的選擇性并縮短了檢測時間。